Niche selection in bacterioplankton: A study of taxonomic composition and single-cell characteristics in an acidic reservoir.

Niche selection and microbial dispersal are key factors that shape microbial communities. However, their relative significance varies across different environments and spatiotemporal scales. While most studies focus on the impact of these forces on community composition, few consider other structural levels such as the physiological stage of the microbial community and single-cell characteristics. To understand the relative influence of microbial dispersal and niche selection on various community structural levels, we concurrently examined the taxonomic composition, abundance and single-cell characteristics of bacterioplankton in an acidic reservoir (El Sancho, Spain) during stratification and mixing periods. A cluster analysis based on environmental variables identified five niches during stratification and one during mixing. Canonical correspondence analysis (CCA) revealed that communities within each niche differed in both, taxonomic and single-cell characteristics. The environmental variables that explained the variation in class-based ordination differed from those explaining the ordination based on single-cell characteristics. However, a Procrustes analysis indicated a high correlation between the CCA ordinations based on both structural levels, suggesting simultaneous changes in the microbial community at multiple structural levels. Our findings underscore the dominant role of environmental selection in occupying different microbial niches, given that microbial dispersal was not restricted.

Trophic status of a coastal lagoon - marine harbor system: Potential outwelling rates to the Mesoamerican Barrier Reef southern region.

Eutrophication is still a serious problem in many coastal areas, including the tropics, where river discharges of nutrients is usually high. The ecological stability and ecosystem services of the Mesoamerican Barrier Reef System (MBRS), the world's second-largest coral reef system, suffer a generalized impact by riverine discharge of sediment and organic and inorganic nutrients, which may lead to coastal eutrophication and a coral-macroalgal phase shift. However, few data exist on the MRBS coastal zone status, particularly in Honduras. Here, two in situ sampling campaigns were carried out (May 2017 and January 2018) in the Alvarado Lagoon and Puerto Cortés Bay (Honduras). Measurements included water column nutrients, chlorophyll-a (Chla), particulate organic and inorganic matter and net community metabolism, completed with satellite images analysis. The lagoon and bay environments are ecologically different systems and present different sensitivities to seasonal changes in precipitation as shown by the multivariate analysis. Nonetheless, net community production and respiration rates were neither different spatially, nor seasonally. In addition, both environments were highly eutrophic as shown by the TRIX index. Thus, the Puerto Cortés system represents an important source of dissolved nutrients and particulate matter to the coastal zone. Even though offshore, water quality, based on estimated outwelling rates from the Puerto Cortés system to the coastal waters of the southern MRBS region, improved considerably, concentrations of Chla and nutrients remained higher than those typically measured in non-polluted coral reefs in the Caribbean region and the suggested threshold values. In situ monitoring and assessment of these aspects are crucial to evaluate the ecological functioning of and threats on the MBRS, and elaborate and implement adequate policies for integrated management given its regional and global importance.

Seasonal cycles of phytoplankton biomass and primary production in a tropical temporarily open-closed estuarine lagoon - The effect of an extreme climatic event.

Temporarily open-closed estuaries and estuarine lagoons are among the most complex aquatic ecosystems, prone to undergo rapid changes in response to global change and other anthropogenic impacts. Nonetheless, studies on the factors that control annual cycles of phytoplanktonic biomass and primary production in such systems, especially tropical ones, are still scarce. Even less information exists on the effect increasingly frequent extreme climatic events (ECE) might have on their dynamics. For this purpose, we monitored the changes in ecological conditions in the Los Micos estuarine lagoon (Honduras) by sampling monthly during an annual cycle that included several changes in the lagoon's mouth phase and attempted to understand which environmental factors affect phytoplanktonic biomass and primary production. We also evaluated the impact of, and recovery from, a tropical storm ECE. Annual mean net production (Pn), integrated for the euphotic zone, (4.3 ± 2.8 gC m-2 d-1) and Chlorophyll a (27.1 ± 19.1 mg m-3) values in Los Micos place it as one of the more productive estuaries worldwide. The physico-chemical characteristics of the lagoon clearly depended on mouth phase; however, the values of Chla and Pn did not show significant differences between the open and closed phases. The application of distance-based multivariate linear models did not show any clear dominant model being able to explain the observed Chla and Pn patterns. The most parsimonious models included among others, salinity, particulate organic carbon and PO43-, which suggests that primary production is controlled by multiple factors. During the ECE, about 19% of DIN, 91% of DSi and PO43-, 60% of particulate organic carbon and nitrogen, and 86% of Chla were exported to the sea, greatly reducing Pn. However, Chla and Pn values recovered to pre-storm levels within 30 days, indicating that these biological variables are highly resilient in Los Micos Lagoon.

Tidal elevation is the key factor modulating burial rates and composition of organic matter in a coastal wetland with multiple habitats.

This study examines long-term burial rates of organic carbon (OC), organic nitrogen (ON), and total sulphur (TS) in a tidal-dominated coastal wetland with a high spatial heterogeneity and habitat diversity, and long history of human impacts, Cádiz Bay (SW Spain). Using replicate sediment cores, we quantified fluxes of these elements over a transect, extending from the lower saltmarsh (Spartina maritima, ~0.3 m mean sea level, MSL) to the lower intertidal region (Zostera noltei, ~ - 0.7 m MSL). Potential organic matter (OM) sources to the sediment were examined using an extensive dataset on carbon and nitrogen stable isotopes, and C:N molar ratios of primary producers in the region. OC burial rates decreased from the sites below MSL (~80 gC·m-2·y-1) to the lower saltmarsh (~50 gC·m-2·y-1), whereas ON burial rates showed an opposite pattern (~3 gN·m-2·y-1 and ~4 gN·m-2·y-1 observed below and above MSL, respectively). TS burial rates (0.5-46 gS·m-2·y-1) did not show any trend along the sea-land gradient. Hence, (tidal) elevation appeared to be an important determinant of sediment biogeochemical properties, and predictor of OM burial rates. The Bayesian mixing model suggested a well-mixed combination of subtidal and terrestrial/high-marsh OM sources to the surface sediments, with no clear indication of an increased contribution from the particular vegetation species inhabiting the sediments. The indication that there is substantial transport, remineralization and cycling of OM between habitats, suggests diversity may play an important role in maintaining this function, reinforcing the idea that a holistic, catchment-scale view is appropriate for understanding and preserving the long-term burial of OM in coastal wetlands.

Water column dissolved silica concentration limits microphytobenthic primary production in intertidal sediments.

Primary production of microphytobenthos (MPB) contributes significantly to the total production in shallow coastal environments. MPB is a diverse community in which diatoms are usually the main microalgal group. Diatoms require N, P, and other nutrients as with other autotrophs, but in addition require silicate to create their outer cell wall. Therefore, dissolved silica (DSi) might be a potential limiting factor for benthic primary production in areas with reduced freshwater input. To test this hypothesis, a microcosm experiment was conducted using intact sediment cores collected from an intertidal mudflat in the Bay of Cádiz and supplied with increasing concentrations of DSi (0, 5, 10, 25, and 45 µmol · L-1 ). After 7 d of enrichment, we determined chlorophyll a and c (Chl a, c) contents, metabolic rates (Net [Pn ] and Areal Gross [PgA ] Production and Light [RL ] and Dark [RD ] Respiration), as well as fluxes of inorganic nutrients across the sediment-water interface. Chl a and c contents increased significantly with respect to the initial conditions but no differences between treatments were found. Both Pn and PgA showed a saturating-like pattern with silicate concentration, reaching maximum rates at a DSi concentration of 45 µmol · L-1 . The addition of DSi also resulted in an increase of DSi and ammonium uptake by the sediment, which was significantly higher in light than in darkness. Our results clearly show that water column DSi concentrations have a direct impact on benthic primary production, also controlling other related processes such as inorganic nutrient fluxes.

Different Types of Diatom-Derived Extracellular Polymeric Substances Drive Changes in Heterotrophic Bacterial Communities from Intertidal Sediments.

Intertidal areas support extensive diatom-rich biofilms. Such microphytobenthic (MPB) diatoms exude large quantities of extracellular polymeric substances (EPS) comprising polysaccharides, glycoproteins and other biopolymers, which represent a substantial carbon pool. However, degradation rates of different EPS components, and how they shape heterotrophic communities in sediments, are not well understood. An aerobic mudflat-sediment slurry experiment was performed in the dark with two different EPS carbon sources from a diatom-dominated biofilm: colloidal EPS (cEPS) and the more complex hot-bicarbonate-extracted EPS. Degradation rate constants determined over 9 days for three sediment fractions [dissolved organic carbon (DOC), total carbohydrates (TCHO), and (cEPS)] were generally higher in the colloidal-EPS slurries (0.105-0.123 d-1) compared with the hot-bicarbonate-extracted-EPS slurries (0.060-0.096 d-1). Addition of hot-bicarbonate-EPS resulted in large increases in dissolved nitrogen and phosphorous by the end of the experiment, indicating that the more complex EPS is an important source of regenerated inorganic nutrients. Microbial biomass increased ~4-6-fold over 9 days, and pyrosequencing of bacterial 16S rRNA genes revealed that the addition of both types of EPS greatly altered the bacterial community composition (from 0 to 9 days) compared to a control with no added EPS. Bacteroidetes (especially Tenacibaculum) and Verrucomicrobia increased significantly in relative abundance in both the hot-bicarbonate-EPS and colloidal-EPS treatments. These differential effects of EPS fractions on carbon-loss rates, nutrient regeneration and microbial community assembly improve our understanding of coastal-sediment carbon cycling and demonstrate the importance of diverse microbiota in processing this abundant pool of organic carbon.

Dynamics of Inorganic Nutrients in Intertidal Sediments: Porewater, Exchangeable, and Intracellular Pools.

The study of inorganic nutrients dynamics in shallow sediments usually focuses on two main pools: porewater (PW) nutrients and exchangeable (EX) ammonium and phosphate. Recently, it has been found that microphytobenthos (MPB) and other microorganisms can accumulate large amounts of nutrients intracellularly (IC), highlighting the biogeochemical importance of this nutrient pool. Storing nutrients could support the growth of autotrophs when nutrients are not available, and could also provide alternative electron acceptors for dissimilatory processes such as nitrate reduction. Here, we studied the magnitude and relative importance of these three nutrient pools (PW, IC, and EX) and their relation to chlorophylls (used as a proxy for MPB abundance) and organic matter (OM) contents in an intertidal mudflat of Cadiz Bay (Spain). MPB was localized in the first 4 mm of the sediment and showed a clear seasonal pattern; highest chlorophylls content was found during autumn and lowest during spring-summer. The temporal and spatial distribution of nutrients pools and MPB were largely correlated. Ammonium was higher in the IC and EX fractions, representing on average 59 and 37% of the total ammonium pool, respectively. Similarly, phosphate in the IC and EX fractions accounted on average for 40 and 31% of the total phosphate pool, respectively. Nitrate in the PW was low, suggesting low nitrification activity and rapid consumption. Nitrate accumulated in the IC pool during periods of moderate MPB abundance, being up to 66% of the total nitrate pool, whereas it decreased when chlorophyll concentration peaked likely due to a high nitrogen demand. EX-Nitrate accounted for the largest fraction of total sediment nitrate, 66% on average. The distribution of EX-Nitrate was significantly correlated with chlorophyll and OM, which probably indicates a relation of this pool to an increased availability of sites for ionic adsorption. This EX-Nitrate pool could represent an alternative nitrate source with significant concentrations available to the microbial community, deeper in the sediment below the oxic layer.

Kinetics of Indigenous Nitrate Reducing Sulfide Oxidizing Activity in Microaerophilic Wastewater Biofilms.

Nitrate decreases sulfide release in wastewater treatment plants (WWTP), but little is known on how it affects the microzonation and kinetics of related microbial processes within the biofilm. The effect of nitrate addition on these properties for sulfate reduction, sulfide oxidation, and oxygen respiration were studied with the use of microelectrodes in microaerophilic wastewater biofilms. Mass balance calaculations and community composition analysis were also performed. At basal WWTP conditions, the biofilm presented a double-layer system. The upper microaerophilic layer (~300 µm) showed low sulfide production (0.31 µmol cm-3 h-1) and oxygen consumption rates (0.01 µmol cm-3 h-1). The anoxic lower layer showed high sulfide production (2.7 µmol cm-3 h-1). Nitrate addition decreased net sulfide production rates, caused by an increase in sulfide oxidation rates (SOR) in the upper layer, rather than an inhibition of sulfate reducing bacteria (SRB). This suggests that the indigenous nitrate reducing-sulfide oxidizing bacteria (NR-SOB) were immediately activated by nitrate. The functional vertical structure of the biofilm changed to a triple-layer system, where the previously upper sulfide-producing layer in the absence of nitrate split into two new layers: 1) an upper sulfide-consuming layer, whose thickness is probably determined by the nitrate penetration depth within the biofilm, and 2) a middle layer producing sulfide at an even higher rate than in the absence of nitrate in some cases. Below these layers, the lower net sulfide-producing layer remained unaffected. Net SOR varied from 0.05 to 0.72 µmol cm-3 h-1 depending on nitrate and sulfate availability. Addition of low nitrate concentrations likely increased sulfate availability within the biofilm and resulted in an increase of both net sulfate reduction and net sulfide oxidation by overcoming sulfate diffusional limitation from the water phase and the strong coupling between SRB and NR-SOB syntrophic relationship.

Photosynthetic activity and community shifts of microphytobenthos covered by green macroalgae.

Macroalgae blooms, a frequent consequence of eutrophication in coastal areas, affect the photosynthetic activity of sediments dominated by microphytobenthos (MPB). Light spectra, steady-state (after 1 h) microprofiles of O2 , gross photosynthesis (Pg ), community respiration in light (RL ) and net community photosynthesis (Pn ) were measured in diatom- and cyanobacteria-dominated communities below increasing layers of Ulva. Photosynthetic photon flux (PPF) decreased exponentially with increasing layers of algae and the light spectrum was increasingly enriched in the green and deprived in blue and red regions. Sediment Pg , Pn and RL decreased as the number of Ulva layers increased; however, 1.6 times higher macroalgal density was necessary to fully inhibit cyanobacteria Pg compared with diatoms, indicating that cyanobacteria were better adapted to this light environment. Long-term (3 weeks) incubations of diatom-dominated sediments below increasing layers of Ulva resulted in a shift in the taxonomic composition of the MPB towards cyanobacteria. Hence, changes in the light climate below macroalgal accumulations can negatively affect the photosynthetic activity of sediments. However, spectral niche differentiation of MPB taxonomic groups and concurrent changes in the MPB community may provide sediments with increased resilience to the detrimental effects of eutrophication.

Nitrate stimulation of indigenous nitrate-reducing, sulfide-oxidising bacterial community in wastewater anaerobic biofilms.

The role of the nitrate-reducing, sulfide-oxidising bacteria (NR-SOB) in the nitrate-mediated inhibition of sulfide net production by anaerobic wastewater biofilms was analyzed in two experimental bioreactors, continuously fed with the primary effluent of a wastewater treatment plant, one used as control (BRC) and the other one supplemented with nitrate (BRN). This study integrated information from H(2)S and pH microelectrodes, RNA-based molecular techniques, and the time course of biofilm growth and bioreactors water phase. Biofilms were a net source of sulfide for the water phase (2.01 micromol S(2-)(tot)m(-2)s(-1)) in the absence of nitrate dosing. Nitrate addition effectively led to the cessation of sulfide release from biofilms despite which a low rate of net sulfate reduction activity (0.26 micromol S(2-)(tot)m(-2)s(-1)) persisted at a deep layer within the biofilm. Indigenous NR-SOB including Thiomicrospira denitrificans, Arcobacter sp., and Thiobacillus denitrificans were stimulated by nitrate addition resulting in the elimination of most sulfide from the biofilms. Active sulfate reducing bacteria (SRB) represented comparable fractions of total metabolically active bacteria in the libraries obtained from BRN and BRC. However, we detected changes in the taxonomic composition of the SRB community suggesting its adaptation to a higher level of NR-SOB activity in the presence of nitrate.

Nitrate promotes biological oxidation of sulfide in wastewaters: experiment at plant-scale.

Biogenic production of sulfide in wastewater treatment plants involves odors, toxicity and corrosion problems. The production of sulfide is a consequence of bacterial activity, mainly sulfate-reducing bacteria (SRB). To prevent this production, the efficiency of nitrate addition to wastewater was tested at plant-scale by dosing concentrated calcium nitrate (Nutriox) in the works inlet. Nutriox dosing resulted in a sharp decrease of sulfide, both in the air and in the bulk water, reaching maximum decreases of 98.7% and 94.7%, respectively. Quantitative molecular microbiology techniques indicated that the involved mechanism is the development of the nitrate-reducing, sulfide-oxidizing bacterium Thiomicrospira denitrificans instead of the direct inhibition of the SRB community. Denitrification rate in primary sedimentation tanks was enhanced by nitrate, being this almost completely consumed. No significant increase of inorganic nitrogen was found in the discharged effluent, thus reducing potential environmental hazards to receiving waters. This study demonstrates the effectiveness of nitrate addition in controlling sulfide generation at plant-scale, provides the mechanism and supports the environmental adequacy of this strategy.